BACKGROUND Cancer detection is a global research focus,and novel,rapid,and label-free techniques are being developed for routine clinical practice.This has led to the development of new tools and techniques from the b...BACKGROUND Cancer detection is a global research focus,and novel,rapid,and label-free techniques are being developed for routine clinical practice.This has led to the development of new tools and techniques from the bench side to routine clinical practice.In this study,we present a method that uses Raman spectroscopy(RS)to detect cancer in unstained formalin-fixed,resected specimens of the esophagus and stomach.Our method can record a clear Raman-scattered light spectrum in these specimens,confirming that the Raman-scattered light spectrum changes because of the histological differences in the mucosal tissue.AIM To evaluate the use of Raman-scattered light spectrum for detecting endoscopically resected specimens of esophageal squamous cell carcinoma(SCC)and gastric adenocarcinoma(AC).METHODS We created a Raman device that is suitable for observing living tissues,and attempted to acquire Raman-scattered light spectra in endoscopically resected specimens of six esophageal tissues and 12 gastric tissues.We evaluated formalin-fixed tissues using this technique and captured shifts at multiple locations based on feasibility,ranging from six to 19 locations 200 microns apart in the vertical and horizontal directions.Furthermore,a correlation between the obtained Raman scattered light spectra and histopathological diagnosis was performed.RESULTS We successfully obtained Raman scattered light spectra from all six esophageal and 12 gastric specimens.After data capture,the tissue specimens were sent for histopathological analysis for further processing because RS is a label-free methodology that does not cause tissue destruction or alterations.Based on data analysis of molecular-level substrates,we established cut-off values for the diagnosis of esophageal SCC and gastric AC.By analyzing specific Raman shifts,we developed an algorithm to identify the range of esophageal SCC and gastric AC with an accuracy close to that of histopathological diagnoses.CONCLUSION Our technique provides qualitative information for real-time morphological diagnosis.However,further in vivo evaluations require an excitation light source with low human toxicity and large amounts of data for validation.展开更多
Stimulated Raman scattering(SRS)microscopy is a highly sensitive chemical imaging technique.However,the SRS imaging performance hinges on two key factors:the reliance on low-noise but bulky solid-state laser sources a...Stimulated Raman scattering(SRS)microscopy is a highly sensitive chemical imaging technique.However,the SRS imaging performance hinges on two key factors:the reliance on low-noise but bulky solid-state laser sources and stringent sample requirements necessitated by high numerical aperture(NA)optics.Here,we present a fiber laser based stimulated Raman photothermal(SRP)microscope that addresses these limitations.While appreciating the portability and compactness of a noisy source,fiber laser SRP enables a two-order-of-magnitude improvement in signal to noise ratio over fiber laser SRS without balance detection.Furthermore,with the use of low NA,long working distance optics for signal collection,SRP expands the allowed sample space from millimeters to centimeters,which diversifies the sample formats to multiwell plates and thick tissues.The sensitivity and imaging depth are further amplified by using urea for both thermal enhancement and tissue clearance.Together,fiber laser SRP microscopy provides a robust,user-friendly platform for diverse applications.展开更多
We report tensorial tomographic Fourier ptychography(T^(2)oFu),a nonscanning label-free tomographic microscopy method for simultaneous imaging of quantitative phase and anisotropic specimen information in 3D.Built upo...We report tensorial tomographic Fourier ptychography(T^(2)oFu),a nonscanning label-free tomographic microscopy method for simultaneous imaging of quantitative phase and anisotropic specimen information in 3D.Built upon Fourier ptychography,a quantitative phase imaging technique,T^(2)oFu additionally highlights the vectorial nature of light.The imaging setup consists of a standard microscope equipped with an LED matrix,a polarization generator,and a polarization-sensitive camera.Permittivity tensors of anisotropic samples are computationally recovered from polarized intensity measurements across three dimensions.We demonstrate T^(2)oFu’s efficiency through volumetric reconstructions of refractive index,birefringence,and orientation for various validation samples,as well as tissue samples from muscle fibers and diseased heart tissue.Our reconstructions of healthy muscle fibers reveal their 3D fine-filament structures with consistent orientations.Additionally,we demonstrate reconstructions of a heart tissue sample that carries important polarization information for detecting cardiac amyloidosis.展开更多
基金Supported by MEXT KAKENHI,JP17K09022 and JP20K07643.
文摘BACKGROUND Cancer detection is a global research focus,and novel,rapid,and label-free techniques are being developed for routine clinical practice.This has led to the development of new tools and techniques from the bench side to routine clinical practice.In this study,we present a method that uses Raman spectroscopy(RS)to detect cancer in unstained formalin-fixed,resected specimens of the esophagus and stomach.Our method can record a clear Raman-scattered light spectrum in these specimens,confirming that the Raman-scattered light spectrum changes because of the histological differences in the mucosal tissue.AIM To evaluate the use of Raman-scattered light spectrum for detecting endoscopically resected specimens of esophageal squamous cell carcinoma(SCC)and gastric adenocarcinoma(AC).METHODS We created a Raman device that is suitable for observing living tissues,and attempted to acquire Raman-scattered light spectra in endoscopically resected specimens of six esophageal tissues and 12 gastric tissues.We evaluated formalin-fixed tissues using this technique and captured shifts at multiple locations based on feasibility,ranging from six to 19 locations 200 microns apart in the vertical and horizontal directions.Furthermore,a correlation between the obtained Raman scattered light spectra and histopathological diagnosis was performed.RESULTS We successfully obtained Raman scattered light spectra from all six esophageal and 12 gastric specimens.After data capture,the tissue specimens were sent for histopathological analysis for further processing because RS is a label-free methodology that does not cause tissue destruction or alterations.Based on data analysis of molecular-level substrates,we established cut-off values for the diagnosis of esophageal SCC and gastric AC.By analyzing specific Raman shifts,we developed an algorithm to identify the range of esophageal SCC and gastric AC with an accuracy close to that of histopathological diagnoses.CONCLUSION Our technique provides qualitative information for real-time morphological diagnosis.However,further in vivo evaluations require an excitation light source with low human toxicity and large amounts of data for validation.
基金supported by NIH grants R35GM136223, R01EB032391, R01EB035429 to JXC.
文摘Stimulated Raman scattering(SRS)microscopy is a highly sensitive chemical imaging technique.However,the SRS imaging performance hinges on two key factors:the reliance on low-noise but bulky solid-state laser sources and stringent sample requirements necessitated by high numerical aperture(NA)optics.Here,we present a fiber laser based stimulated Raman photothermal(SRP)microscope that addresses these limitations.While appreciating the portability and compactness of a noisy source,fiber laser SRP enables a two-order-of-magnitude improvement in signal to noise ratio over fiber laser SRS without balance detection.Furthermore,with the use of low NA,long working distance optics for signal collection,SRP expands the allowed sample space from millimeters to centimeters,which diversifies the sample formats to multiwell plates and thick tissues.The sensitivity and imaging depth are further amplified by using urea for both thermal enhancement and tissue clearance.Together,fiber laser SRP microscopy provides a robust,user-friendly platform for diverse applications.
基金a Duke-Coulter Translational Partnership and funding from a 3M Nontenured Faculty Awarda grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute(KHIDI),funded by the Ministry of Health&Welfare,Republic of Korea(Grant No.HI19C1344)funding from the German academic exchange service(DAAD,project 57698081)
文摘We report tensorial tomographic Fourier ptychography(T^(2)oFu),a nonscanning label-free tomographic microscopy method for simultaneous imaging of quantitative phase and anisotropic specimen information in 3D.Built upon Fourier ptychography,a quantitative phase imaging technique,T^(2)oFu additionally highlights the vectorial nature of light.The imaging setup consists of a standard microscope equipped with an LED matrix,a polarization generator,and a polarization-sensitive camera.Permittivity tensors of anisotropic samples are computationally recovered from polarized intensity measurements across three dimensions.We demonstrate T^(2)oFu’s efficiency through volumetric reconstructions of refractive index,birefringence,and orientation for various validation samples,as well as tissue samples from muscle fibers and diseased heart tissue.Our reconstructions of healthy muscle fibers reveal their 3D fine-filament structures with consistent orientations.Additionally,we demonstrate reconstructions of a heart tissue sample that carries important polarization information for detecting cardiac amyloidosis.
